Nucleic Acid Amplification Technique for Rapid Diagnostic Analysis


CDC researchers developed a simple target-specific isothermal nucleic acid amplification technique, termed Genome Exponential Amplification Reaction (GEAR). The method employs a set of four primers (two inner and two outer). The outer primer pair targets three specific nucleic acid sequences at a constant 60°C, while the inner pair of primers accelerates and improves the sensitivity of the assay. The GEAR technique is an improvement over loop-mediated isothermal amplification (LAMP) in three ways. First, the GEAR method uses two Tab primers which target three genomic regions (corresponding LAMP primers target four regions). Second, the GEAR method features complementary 5' ends between the forward and reverse primers. Third, the GEAR method does not require a second set of outer primers (LAMP requires two outermost primers). Additionally, the GEAR isothermal method can be performed in a relatively inexpensive water bath or heating block, with detection of amplification products by fluorescence, thus making it suitable for low resource settings.

Potential Commercial Applications: Competitive Advantages:
  • Rapid diagnostic analysis of biological samples
  • Qualitative and quantitative analysis of nucleic acids
  • Low-cost diagnostics for malaria, tuberculosis, and other infectious diseases
 
  • Rapid, portable, cost-effective
  • Useful in low resource settings
  • A “single-tube” assay that eliminates need for thermal cyclers or gel electrophoresis
  • Unlike many other isothermal amplification approaches, GEAR can be efficiently performed at temperatures exceeding 60 °C, increasing specificity and accuracy


Related Invention(s):



Inventors:
Jothikumar Narayanan (CDC)
Prithiviraj Jothikumar (CDC)
Vincent Hill (CDC)


Intellectual Property:
PCT Application No. PCT/US2012/049784


Publications:
Prithiviraj J, et al. PMID 22450319
Jothikumar P, et al. PMID 24424127

Licensing Contact:
Whitney Blair ,
NIH Office of Technology Transfer
6011 Executive Blvd. Suite 325
Rockville , MD 20852
Email: blairw2@mail.nih.gov
Phone: 301-435-4937
Fax: 301-402-0220

OTT Reference No: E-273-2013/0

Updated: 02/10/2014